Development and validation of UPLC-MS/MS method for in vitro quantitative analysis of pyrazinamide in lipid core-shell nanoarchitectonics for improved metabolic stability

AbstractPyrazinamide (PZA), a medication for tuberculosis, has high aqueous solubility and low permeability, undergoes extensive liver metabolism, and exhibits liver toxicity through its metabolites. To avoid this, PZA in lipid core-shell nanoarchitectonics has been formulated to target lymphatic uptake and provide metabolic stability to the incorporated drug. The UPLC-MS/MS method for reliable in vitro quantitative analysis of pyrazinamide (PZA) in lipid core-shell nanoarchitectonics as per ICH guidance was developed and validated using the HILIC column. The developed UPLC-MS/MS method is a simple, precise, accurate, reproducible, and sensitive method for the estimation of PZA in PZA-loaded lipid core-shell nanoarchitectonics for the in vitro determination of % entrapment efficiency, % loading of pyrazinamide, and microsomal stability of lipid core-shell nanoarchitectonics in human liver microsomes. The % entrapment efficiency was found to be 42.72% (±12.60). Lipid nanoarchitectonics was found to be stable in human liver microsomes, where %QH was found to be 6.20%, that is, low clearance. Thus, this formulation is suitable for preventing PZA-mediated extensive liver metabolism. These findings are relevant for the development of other lipid-mediated, suitable, stable nanoformulations containing PZA through various in vitro methods.

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Monocarbonyl curcuminoids as antituberculosis agents with their moderate in‐vitro metabolic stability on human liver microsomes

Journal of Biochemical and Molecular Toxicology ◽

10.1002/jbt.22754 ◽

2021 ◽

Author(s):

Gagandeep ◽

Manisha Singh ◽

Saqib Kidawi ◽

Ujjalkumar S. Das ◽

Thirumurthy Velpandian ◽

...

Keyword(s):

Human Liver ◽

Liver Microsomes ◽

Human Liver Microsomes ◽

Metabolic Stability

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In vitro metabolic stability of moisture-sensitive rabeprazole in human liver microsomes and its modulation by pharmaceutical excipients

Archives of Pharmacal Research ◽

10.1007/s12272-001-1171-z ◽

2008 ◽

Vol 31 (3) ◽

pp. 406-413 ◽

Cited By ~ 12

Author(s):

Shan Ren ◽

Mi-Jin Park ◽

Aera Kim ◽

Beom-Jin Lee

Keyword(s):

Human Liver ◽

Liver Microsomes ◽

Human Liver Microsomes ◽

Metabolic Stability ◽

Pharmaceutical Excipients ◽

Moisture Sensitive

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Evaluation of Diallyl Sulfide Analogs for Cytotoxicity, Inhibition of CYP2E1, and Metabolite Profiling Using In Vitro Cell Culture, Human Liver Microsomes and LCMS/MS

10.26226/morressier.57d6b2b8d462b8028d88ee12 ◽

2016 ◽

Author(s):

Mohammad Rahman

Keyword(s):

Cell Culture ◽

Metabolite Profiling ◽

Human Liver ◽

Liver Microsomes ◽

Human Liver Microsomes ◽

Diallyl Sulfide ◽

In Vitro Cell Culture

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Nontargeted Metabolomics by High-Resolution Mass Spectrometry to Study the In Vitro Metabolism of a Dual Inverse Agonist of Estrogen-Related Receptors β and γ, DN203368

Pharmaceutics ◽

10.3390/pharmaceutics13060776 ◽

2021 ◽

Vol 13 (6) ◽

pp. 776

Author(s):

Sin-Eun Kim ◽

Seung-Bae Ji ◽

Euihyeon Kim ◽

Minseon Jeong ◽

Jina Kim ◽

...

Keyword(s):

Mass Spectrometry ◽

High Resolution ◽

Human Liver ◽

High Resolution Mass Spectrometry ◽

Liver Microsomes ◽

Human Liver Microsomes ◽

Rat Liver Microsomes ◽

High Resolution Mass ◽

Resolution Mass

DN203368 ((E)-3-[1-(4-[4-isopropylpiperazine-1-yl]phenyl) 3-methyl-2-phenylbut-1-en-1-yl] phenol) is a 4-hydroxy tamoxifen analog that is a dual inverse agonist of estrogen-related receptor β/γ (ERRβ/γ). ERRγ is an orphan nuclear receptor that plays an important role in development and homeostasis and holds potential as a novel therapeutic target in metabolic diseases such as diabetes mellitus, obesity, and cancer. ERRβ is also one of the orphan nuclear receptors critical for many biological processes, such as development. We investigated the in vitro metabolism of DN203368 by conventional and metabolomic approaches using high-resolution mass spectrometry. The compound (100 μM) was incubated with rat and human liver microsomes in the presence of NADPH. In the metabolomic approach, the m/z value and retention time information obtained from the sample and heat-inactivated control group were statistically evaluated using principal component analysis and orthogonal partial least-squares discriminant analysis. Significant features responsible for group separation were then identified using tandem mass spectra. Seven metabolites of DN203368 were identified in rat liver microsomes and the metabolic pathways include hydroxylation (M1-3), N-oxidation (M4), N-deisopropylation (M5), N,N-dealkylation (M6), and oxidation and dehydrogenation (M7). Only five metabolites (M2, M3, and M5-M7) were detected in human liver microsomes. In the conventional approach using extracted ion monitoring for values of mass increase or decrease by known metabolic reactions, only five metabolites (M1-M5) were found in rat liver microsomes, whereas three metabolites (M2, M3, and M5) were found in human liver microsomes. This study revealed that nontargeted metabolomics combined with high-resolution mass spectrometry and multivariate analysis could be a more efficient tool for drug metabolite identification than the conventional approach. These results might also be useful for understanding the pharmacokinetics and metabolism of DN203368 in animals and humans.

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